Targeted inCITE-Seq Analysis Identifies the Loss of Nuclear TDP-43 in Endothelium as a Mediator of Blood Brain Barrier Signaling Pathway Dysfunction in Neurodegeneration.
Targeted inCITE-Seq Analysis Identifies the Loss of Nuclear TDP-43 in Endothelium as a Mediator of Blood Brain Barrier Signaling Pathway Dysfunction in Neurodegeneration.
复制标题
靶向 inCITE-Seq 分析确定内皮细胞中核 TDP-43 的丢失是神经退行性变中血脑屏障信号通路功能障碍的介质。
DOI：
10.1101/2023.12.13.571178
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发表时间：
2023
期刊：
影响因子：
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通讯作者：
Murphy,PatrickA
中科院分区：
文献类型：
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作者：
Omar,OmarMF;Kimble,AmyL;Cheemala,Ashok;Tyburski,JordanD;Pandey,Swati;Wu,Qian;Reese,Bo;Jellison,EvanR;Li,Yunfeng;Hao,Bing;Yan,Riqiang;Murphy,PatrickA
尽管内皮细胞在衰老和神经退行性疾病中对血脑屏障(BBB)的调节非常重要，但提取内皮细胞(EC)核的困难限制了对这些细胞的分析。此外，几乎所有肌萎缩侧索硬化症(ALS)和额颞部变性(FTD)以及很大一部分阿尔茨海默病(AD)都表现出神经元TDP-43聚集，导致核功能丧失，但TDP-43在人血脑屏障内皮细胞中是否也有类似的变化尚不清楚。在这里，我们利用一种新的技术，从人的皮质脑组织中浓缩内皮和小胶质细胞核，结合INSTE-SEQ，分析大量健康和疾病捐赠者的核蛋白和RNA转录本。我们的发现揭示了在神经退行性变状态下，近一半的毛细血管内皮细胞具有独特的转录特征，其特征是核β-连环蛋白和标准下游基因水平降低，而肿瘤坏死因子/核因子-kB靶基因增加。我们证明，这与核p65/核因子-kB的增加无关，而是与这些疾病相关的内皮细胞中核TDP-43的特异性丢失有关。在靶向干扰TDP-43的动物模型中的比较分析表明，这足以驱动这些转录改变。这项工作揭示了TDP-43是核p65/NF-kB转录输出的关键调控因子，它在屏障维持和屏障妥协炎症反应中具有矛盾的作用，并表明在AD、ALS和FTD的进展中观察到的BBB缺陷与ECs中的疾病特异性丢失有关。
Despite the importance of the endothelium in the regulation of the blood brain barrier (BBB) in aging and neurodegenerative disease, difficulties in extracting endothelial cell (EC) nuclei have limited analysis of these cells. In addition, nearly all Amyotrophic Lateral Sclerosis (ALS) and Frontotemporal Degeneration (FTD), and a large portion of Alzheimer’s Disease (AD) exhibit neuronal TDP-43 aggregation, leading to loss of nuclear function, but whether TDP-43 is similarly altered in human BBB ECs is unknown. Here we utilize a novel technique for the enrichment of endothelial and microglial nuclei from human cortical brain tissues, combined with inCITE-seq, to analyze nuclear proteins and RNA transcripts in a large cohort of healthy and diseased donors. Our findings reveal a unique transcriptional signature in nearly half of the capillary endothelial cells across neurodegenerative states, characterized by reduced levels of nuclear β-Catenin and canonical downstream genes, and an increase in TNF/NF-kB target genes. We demonstrate that this does not correlate with increased nuclear p65/NF-kB, but rather a specific loss of nuclear TDP-43 in these disease associated ECs. Comparative analysis in animal models with targeted disruption of TDP-43 shows that this is sufficient to drive these transcriptional alterations. This work reveals that TDP-43 is a critical governor of the transcriptional output from nuclear p65/NF-kB, which has paradoxical roles in barrier maintenance and also barrier compromising inflammatory responses, and suggests that disease specific loss in ECs contributes to BBB defects observed in the progression of AD, ALS and FTD.